A small population of HIV-1-infected individuals remains clinically healthy and immunologically normal for more than ten years. We have studied ten subjects who have been asymptomatic with normal and stable CD4+ lymphocyte counts, despite 12 to 16 years of HIV-1 infection, to gain information on the determinants of nonprogression. Multiple methods were used to determine the viral load in their blood. Plasma cultures were uniformly negative for infectious virus. However, particle-associated HIV-1 RNA was detectable in four subjects using a sensitive branched DNA amplification assay. In peripheral blood mononuclear cells (PBMC), infectious HIV-1 was quantified in three subjects using a standard limiting-dilution culture method. Infectious virus was recovered from another subject using a CD8-depleted culture. In contrast, six subjects had no detectable infectious virus in PBMC. All had detectable viral DNA in PBMC by a quantitative polymerase chain reaction assay, but the copy numbers were low, ranging from 10 to 100 copies per 10(6) PBMC in all but two subjects. Overall, the viral burden in the plasma and PBMC of long-term survivors was orders of magnitude lower than those typically found in progressors. Possible mechanisms for low levels of HIV-1 in vivo were examined experimentally.